Next-Generation Sequencing (NGS)

In this case study, we describe a newly developed automated method for the quantification of Illumina NGS libraries using the KAPA Library Quantification Kit on the NGS STAR for Library Prep.

Hamilton Robotics and Illumina, Inc have partnered to automate the TruSight Tumor 170 library preparation kit on the NGS STAR. TruSight Tumor 170 simultaneously analyzes DNA and RNA while covering a wide range of genes and variant types. TruSight Tumor 170 is capable of producing accurate results from low-quality formalin-fixed paraffin-embedded (FFPE) samples with a minimum input requirement of 40 ng. The design of the NGS STAR, as a single configuration, enables implementation of additional methods as well as future Illumina protocols using the TruSight Oncology workflow as they are released.

The BioAnalytica-Genotypos center in Athens chose to couple the Hamilton STARlet robot with the SOPHiA Clinical Exome Solution (CES) for dealing with the increasing workload they were facing. In fact, over the past decade, the number of samples and types of analyses processed in the laboratory increased exponentially, leading to a pressure to develop, optimize, and validate NGS high-throughput assays. To ensure high-quality results on the new workflow and facilitate its implementation, the automated protocol has been extensively tested by SOPHiA GENETICS and Hamilton on a smaller representative panel of 128 genes. The validation study ensured a reliable and flexible automation, which has been efficiently applied on larger gene panels, including CES.

In this application note, we discuss the results and findings from a customer project to automate NEOonsite assays using Hamilton’s NGS STARTM workstation, aiming for a reduction of hands-on time and number of user interventions, while keeping a high level of reproducibility.